LOCUS       sequence-257590-        4529 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Enhancer cloning nanotag reporter vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4529)
  AUTHORS   Williams RM, Candido-Ferreira I, Repapi E, Gavriouchkina D, 
            Senanayake U, Ling ITC, Telenius J, Taylor S, Hughes J, 
            Sauka-Spengler T
  TITLE     Reconstruction of the Global Neural Crest Gene Regulatory Network In
            Vivo.
  JOURNAL   Dev Cell. 2019 Oct 21;51(2):255-276.e7. doi: 
            10.1016/j.devcel.2019.10.003.
  PUBMED    31639368
REFERENCE   2  (bases 1 to 4529)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.devcel.2019.10"; journalName: "Dev Cell"; date: 
            "2019-10-21- 21"; volume: "51"; issue: "2"; pages: "255-276.e7"
COMMENT     Sequence Label: pTK_Cerulean_Nanotag_70_neg_Control
FEATURES             Location/Qualifiers
     source          1..4529
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        65..210
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          288..420
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     403..422
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      474..483
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(480..482,483..485,486..1199)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(525..546)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(786..805)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1133..1154
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1373..1494
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1384..1403)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1438..1457
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1742..1759)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1913..2501)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1993..2012)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2672..3463,3464..3532))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3295..3314
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3533..3637)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3664..4119
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3751..3770)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3961..3982
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4250..4298
                     /note="synthetic polyadenylation signal"
     misc_feature    4312..4403
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4352..4371
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ccgttgctcg cgtttgctgg cggtgtcccc ggaagaaata tatttgcatg tctttagttc
       61 tatgatgaca caaaccccgc ccagcgtctt gtcattggcg aattcgaaca cgcagatgca
      121 gtcggggcgg cgcggtccca ggtccacttc gcatattaag gtgacgcgtg tggcctcgaa
      181 caccgagcga ccctgcagcg acccgcttaa cagcgtcaac agcgtgccgc agatcccaag
      241 cttgcggccg cttaagctgc agaagttggt cgtgaggcac tgggcaggta agtatcaagg
      301 ttacaagaca ggtttaagga gaccaataga aactgggctt gtcgagacag agaagactct
      361 tgcgtttctg ataggcacct attggtctta ctgacatcca ctttgccttt ctctccacag
      421 gtgtccactc ccaggttcaa tacagctctt aagcggccgc aagctagctt gccgccacca
      481 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
      541 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
      601 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
      661 tcgtgaccac cctgacctgg ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc
      721 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
      781 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
      841 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
      901 agctggagta caacgccatc agcgacaacg tctatatcac cgccgacaag cagaagaacg
      961 gcatcaaggc caacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     1021 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     1081 acctgagcac ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     1141 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtgag
     1201 cctctagact ccctcagcgg cgcgccaaca cactggacgc agcctccgca gtgtggtaga
     1261 cgccgtggat atgtcttcgc aaatacggag ccagatcgtg agcagtaagc ttgtctatgg
     1321 tagatctcac gtgtctagag tcggggcggc cggccgcttc gagcagacat gataagatac
     1381 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     1441 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     1501 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     1561 aagtaaaacc tctacaaatg tggtaaaatc gataaggatc cgtcgaccga tgcccttgag
     1621 agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact
     1681 tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tcttccgctt
     1741 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     1801 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     1861 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     1921 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     1981 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2041 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2101 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     2161 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     2221 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     2281 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     2341 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     2401 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     2461 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     2521 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     2581 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     2641 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     2701 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     2761 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     2821 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     2881 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     2941 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     3001 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     3061 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     3121 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     3181 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     3241 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     3301 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     3361 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     3421 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     3481 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     3541 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     3601 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     3661 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     3721 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     3781 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     3841 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     3901 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     3961 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     4021 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     4081 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttg ccattcgcca ttcaggctgc
     4141 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag cccaagctac
     4201 catgataagt aagtaatatt aaggtacggg aggtacttgg agcggccgca ataaaatatc
     4261 tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagtac taacatacgc
     4321 tctccatcaa aacaaaacga aacaaaacaa actagcaaaa taggctgtcc ccagtgcaag
     4381 tgcaggtgcc agaacatttc tctatcgata ggtaccgagc tcgagttcca tgagctggat
     4441 cgatgatatc cgatcgatcg tagcacagga ccgctagccc gggctcgaga tctgcgatct
     4501 aagtaagctt gctgcttcat ccccgtggc
//
